我们所展示的质粒图谱主要是从文献和开放数据库中收集而来，主要是为了方便研究工作，其中一小部分质粒进行了质量控制，可供科学家使用。
所有的产品都严格仅供科学研究使用，不能应用于临床试验，包括人体摄入、注射或外用的药理学用途。
确保质粒的关键元件正确，但是我们并不能保证实验结果。页面展示的图谱序列为理论序列，可能与测序结果不一致，请自行比对后确定是否满足要求。(如果测过序，本页面一般会提供下载)
开放数据库中的大多数载体序列都没有被完全测序。如果实际序列与参考序列的相似度超过99%，则将其视为正确。
由于科学研究是在探索未知，具有很大的不确定性，在任何情况下，我们都不承担超出质粒本身的额外经济损失或责任。
- 载体名称:
- pMCS
- 载体抗性:
- Kanamycin
- 载体长度:
- 12362 bp
- 载体类型:
- Standard reference vector
- 复制子:
- ori
- 载体来源:
- Xi J, Wang X, Dong L, Dong Y, Su Y, Tang Q, Wang Z.
- 启动子:
- CaMV35S(enhanced)
pMCS 载体图谱
质粒操作方法
1. 发货形式：质粒干粉(常温运输，存于-20度，请务必先转化提质粒后使用)
2. 收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O溶解质粒;（质粒复测的浓度有时候与标称值差距较大，这可能是因为冻干质粒在管中的位置、复溶效率、测量偏差以及管壁的吸附导致，因此建议先转化提质粒后再使用）
3. 取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；
4. 加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min (30℃培养1-1.5小时);
5. 6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上;
6. 将平板倒置37℃培养14h，如果要求是30℃则培养20h; (菌落过多则将质粒稀释后再转化；没有菌落则加入10μl质粒转化；建议不要直接转表达感受态， 要先转克隆感受态，重提质粒后再导入表达感受态);
7. 挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
pMCS 载体序列
LOCUS V004804 12362 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004804
VERSION V004804
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12362)
AUTHORS Xi J, Wang X, Dong L, Dong Y, Su Y, Tang Q, Wang Z.
TITLE Construction of a standard reference plasmid containing seven target
genes for the detection of transgenic cotton
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 12362)
AUTHORS Xi J, Wang X, Dong L, Dong Y, Su Y, Tang Q, Wang Z.
TITLE Direct Submission
JOURNAL Submitted (14-APR-2014) Biosafet Lab, Biotechnology Research
Institute, No. 12 Zhongguancun South Street, Beijing 100081, China
REFERENCE 3 (bases 1 to 12362)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12362)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(14-APR-2014) Biosafet Lab, Biotechnology Research Institute, No. 12
Zhongguancun South Street, Beijing 100081, China"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12362
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1219..1845
/label="pVS1 StaA"
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
CDS 2282..3346
/label="pVS1 RepA"
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
rep_origin 3415..3609
/label="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 3953..4093
/label="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(4279..4867)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4957..5748)
/label="KanR"
/note="aminoglycoside phosphotransferase"
misc_feature 6173..6197
/label="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal complement(6275..6449)
/label="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(6509..7297)
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
promoter complement(7366..8043)
/label="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 8234..8255
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8270..8300
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 8308..8324
/label="lac operator"
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 8332..8348
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 8364..8633
/codon_start=1
/gene="cptI"
/product="cowpea trypsin inhibitor"
/label="cptI"
/protein_id="AII71483.1"
/translation="MRMDLKHLGSNHHDDSSDEPSESSEPCCDSCICTKSIPPQCHCTD
IRWNSCHSACKSCMCTRSMPGKCRCLDIADFCYKPCKSRDEDDE"
gene 8364..8633
/gene="cptI"
/label="cptI"
CDS complement(8551..9579)
/codon_start=1
/gene="Cry1Ab/1Ac"
/product="Cry1Ab/Cry1Ac fusion protein"
/label="Cry1Ab/1Ac"
/note="Bt protein"
/protein_id="AII71484.1"
/translation="NCLSNPEVVVLGGERIETGYTPIDISLSLTQFLLSEFVPGAGFVL
GPVDIIWGIFGPSQWDAFLVQIEQLINQRIEEFARNQAISRLEGLSNLYQIYAESFREW
EADPTNPALREEMRIQFNDMNSALTTAIPLFAVQNYQVPLLSVYVQAANLHLSVLRDVS
VFGQRWGFDAATINSRYNDLTRLIGNYTDHAVRWYNTGLERVWGPDSRDWIRYNQFRRE
LTLTVLDIVSLFPNYDSRTYPIRTVSQLTREIYTNPVLENFDGSFRGSAQGIEGSIRSP
HLMDILNSITIYTDAHRGEYYWSGHQIMASPVGFIIRRLRHEICTVCSRSPRCPGNGIY
PA"
gene complement(8551..9579)
/gene="Cry1Ab/1Ac"
/label="Cry1Ab/1Ac"
CDS 9595..10959
/gene="aroA"
/label="3-phosphoshikimate 1-carboxyvinyltransferase"
/note="3-phosphoshikimate 1-carboxyvinyltransferase from
Agrobacterium sp. (strain CP4). Accession#: Q9R4E4"
regulatory 10963..11180
/label="NOS"
/note="NOS"
/regulatory_class="terminator"
CDS join(11182..11296,11671..12064)
/codon_start=1
/gene="SadI"
/product="AE9 stearoyl-ACP desaturase"
/label="SadI"
/note="from cotton K312"
/protein_id="AII71486.1"
/translation="STMPSPRNLRSSLALLFHQRPPLDLPSFP*SPPFLLAPKRLGI*K
SLSRLQRRCLFRSPTPCRLTRLRSLNLWRAGLRTTF*LTSNQLRNVGNPPTFFQILILM
DFMSKSKSLGKGQRRSQMITL*FWLVI*SPRKPFQLIKQCLIPWMELVMRQVLASLALA
VVLQRR"
gene 11182..12064
/gene="SadI"
/label="SadI"
primer_bind complement(12038..12054)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 12257..12281
/label="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"